Epigenetically Modified DNAzyme Walkers for Simultaneously Imaging of MGMT and FTO Activity in Living Cancer Cells

2025 · doi:10.1101/2025.06.15.659742
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Abstract

Investigations into epigenetic modifications have underscored the pivotal roles demethylases play in modulating the dynamics of RNA and DNA. Abnormal demethylase expression is closely related to the occurrence and development of many diseases. Unraveling the individual activity of different demethylases is essential to study the underlying mechanisms. In this paper, we report the development of demethylase-responsive DNAzyme-powered walkers designed for the simultaneous imaging of two important demethylases: O6-methylguanine-DNA methyltransferase (MGMT) and fat mass and obesity-associated protein (FTO). Epigenetically inactivated DNAyzme-based DNA walker-functionalized gold nanoparticles were employed to orthogonally visualize MGMT and FTO activities in various live cancer cell lines with high sensitivity. This method offers a direct way for the comprehensive evaluation of multiple demethylase activities, and the understanding of epigenetic regulation.
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Abstract Investigations into epigenetic modifications have underscored the pivotal roles demethylases play in modulating the dynamics of RNA and DNA. Abnormal demethylase expression is closely related to the occurrence and development of many diseases. Unraveling the individual activity of different demethylases is essential to study the underlying mechanisms. In this paper, we report the development of demethylase-responsive DNAzyme-powered walkers designed for the simultaneous imaging of two important demethylases: O6-methylguanine-DNA methyltransferase (MGMT) and fat mass and obesity-associated protein (FTO). Epigenetically inactivated DNAyzme-based DNA walker-functionalized gold nanoparticles were employed to orthogonally visualize MGMT and FTO activities in various live cancer cell lines with high sensitivity. This method offers a direct way for the comprehensive evaluation of multiple demethylase activities, and the understanding of epigenetic regulation. Competing Interest Statement The authors have declared no competing interest. Footnotes We have updated the title of article and author order based on the actual research contributions. This modification did not alter any content in the main text of the article nor change its key conclusions.

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